U.S. patent application number 17/615196 was filed with the patent office on 2022-07-21 for nutritional composition.
This patent application is currently assigned to MORINAGA MILK INDUSTRY CO., LTD.. The applicant listed for this patent is MORINAGA MILK INDUSTRY CO., LTD.. Invention is credited to Tatsuya Ehara, Yasuaki Wada.
Application Number | 20220226382 17/615196 |
Document ID | / |
Family ID | |
Filed Date | 2022-07-21 |
United States Patent
Application |
20220226382 |
Kind Code |
A1 |
Wada; Yasuaki ; et
al. |
July 21, 2022 |
Nutritional Composition
Abstract
An object of the present invention is to provide a technique for
preventing infection of pathogenic viruses, which cause infectious
diseases in animals or humans. A nutritional composition containing
lactoferrin and/or lactoferrin hydrolysate, and human milk
oligosaccharide is provided. One or more human milk
oligosaccharides are preferably 2'-fucosyllactose,
3-fucosyllactose, 3'-sialyllactose, and 6'-sialyllactose. The
nutritional compositions of the present invention can be used for
preventing infection, especially for preventing viral
infection.
Inventors: |
Wada; Yasuaki; (Kanagawa,
JP) ; Ehara; Tatsuya; (Kanagawa, JP) |
|
Applicant: |
Name |
City |
State |
Country |
Type |
MORINAGA MILK INDUSTRY CO., LTD. |
Tokyo |
|
JP |
|
|
Assignee: |
MORINAGA MILK INDUSTRY CO.,
LTD.
Tokyo
JP
|
Appl. No.: |
17/615196 |
Filed: |
June 5, 2020 |
PCT Filed: |
June 5, 2020 |
PCT NO: |
PCT/JP2020/022294 |
371 Date: |
November 30, 2021 |
International
Class: |
A61K 35/20 20060101
A61K035/20; A61K 38/40 20060101 A61K038/40; A61K 31/702 20060101
A61K031/702; A61K 9/00 20060101 A61K009/00; A23L 33/125 20060101
A23L033/125; A23L 33/19 20060101 A23L033/19; A23L 33/00 20060101
A23L033/00 |
Foreign Application Data
Date |
Code |
Application Number |
Jun 5, 2019 |
JP |
2019-105506 |
Claims
1. A nutritional composition comprising a) lactoferrin, a
lactoferrin hydrolysate, or both; and b) a human milk
oligosaccharide.
2. The nutritional composition of claim 1, wherein mass ratio of
the human milk oligosaccharide to the lactoferrin, lactoferrin
hydrolysate, or both is not lower than 1/100 and not higher than
100.
3. The nutritional composition of claim 1, wherein amount of the
lactoferrin, lactoferrin hydrolysate, or both is 0.001 to 5.0 mass
% based on the entire composition.
4. The nutritional composition of claim 1, wherein the amount of
the human milk oligosaccharide is 0.001 to 10.0 mass % based on the
entire nutritional composition.
5. The nutritional composition of claim 1, wherein the human milk
oligosaccharide is selected from the group consisting of
2'-fucosyllactose, 3-fucosyllactose, 3'-sialyllactose,
6'-sialyllactose, and combinations thereof.
6. A method of preventing infection in a subject, comprising
administering to the subject the nutritional composition of claim
1.
7. A method of preventing a gastrointentional infection in a
subject comprising administering to the subject the nutritional
composition of claim 1.
8. The nutritional composition of claim 1, wherein said composition
is in the form of a powdered formula.
9. The nutritional composition of claim 1, wherein said composition
is in the form of liquid formula.
10. The method of claim 6, wherein said subject is an infant, a
young child, or an elderly person.
11. A method of claim 7, wherein said subject is an infant, a young
child, or an elderly person.
Description
TECHNICAL FIELD
[0001] The present invention relates to a nutritional composition
containing lactoferrin and/or a lactoferrin hydrolysate, and a
human milk oligosaccharide.
BACKGROUND ART
[0002] In nature, there are various microorganisms that cause
infectious diseases in a human or animal. Those microorganisms are
classified into bacteria, mycoplasmas, rickettsias, chlamydias,
fungi, protozoas, and viruses. The genetic material of viruses is
deoxyribonucleic acid (DNA) or ribonucleic acid (RNA), and viruses
can proliferate only by parasitism in host cells, whereas bacteria
and other microorganisms have energy metabolism systems and
heterotrophically proliferate (for example, Non-patent document
1).
[0003] Influenza virus infectious diseases are prevalent in winter,
and infants, young children, elderly persons, and patients with
underlying diseases such as a malignant tumor, heart disease, and
cerebrovascular disease constitute a high-risk group for influenza
virus infection, and often develop life-threatening critical
infections.
[0004] Although antibiotics have been developed and effective
treatments are performed especially for bacterial infectious
diseases among microbial infectious diseases, such antibiotics are
not effective for treating viral infectious diseases. The present
situation is that development of antiviral agents as therapeutic
drugs for viral infectious diseases is not progressing so much even
now in contrast to the rapid progress of chemotherapies using
antibiotics.
[0005] The antiviral agents developed so far include aciclovir,
azidothymidine, and ribavirin, which are nucleic acid analogues
that target replication of viral nucleic acid, zanamivir
(4-guanidinoneuraminic acid), which is a neuraminidase inhibitor,
indinavir, which is a protease inhibitor, and so forth. Although
these antiviral agents as drugs show potent efficacy, side
reactions caused by administration of these cause serious clinical
problems.
[0006] Because of such a situation, there has been continued search
for active ingredients showing antiviral action, which are safe,
show few side reactions, and can be ingested over a long period of
time.
[0007] Lactoferrin (henceforth also abbreviated as LF) is a milk
protein, and has been reported to have various biological functions
such as antibacterial activity, antiviral activity, action for
controlling iron absorption from intestinal tract, cell
proliferating action, and immunoregulating action (Non-patent
document 2), and use thereof as an influenza virus
infection-preventing agent has already been proposed (Patent
document 1). It has also been reported that lactoferrin
decomposition products (peptides derived from lactoferrin) also
show an antiviral activity against influenza viruses and RS viruses
(Patent document 2).
[0008] It has also been reported that human milk oligosaccharide
(henceforth also abbreviated as HMO), which is a general term for
referring to various oligosaccharides contained in human
colostrums, can prevent viral infections (Patent document 3).
PRIOR ART REFERENCES
Patent Documents
[0009] Patent document 1: Japanese Patent No. 3061376
[0010] Patent document 2: Japanese Patent Laid-open (KOKAI) No.
2005-272438
[0011] Patent document 3: Japanese Patent Laid-open (KOHYO) No.
2017-515455
Non-Patent Documents
[0012] Non-patent document 1: Medical Virology, 2nd revised
edition, Ed. by Osato Toyoro, Nankodo, 2000, pp. 15-24
[0013] Non-patent document 2: B. Lonnerdal et al., Ann. Rev. Nutr.,
Vol. 15, 1995, pp. 93-110
SUMMARY OF THE INVENTION
Object to be Achieved by the Invention
[0014] An object of the present invention is to provide a technique
for preventing infection of pathogenic viruses and so forth, which
causes infectious diseases in animals and human.
Means for Achieving the Object
[0015] The inventors of the present invention conducted various
research in order to achieve the aforementioned object, and as a
result, they found that a nutritional composition containing
lactoferrin and/or a lactoferrin hydrolysate, and a human milk
oligosaccharide has an infection-preventing effect, and
accomplished the present invention.
[0016] The present invention thus provides a nutritional
composition containing lactoferrin and/or a lactoferrin
hydrolysate, and a human milk oligosaccharide.
[0017] According to the present invention, the mass ratio of the
human milk oligosaccharide to the lactoferrin and/or lactoferrin
hydrolysate is preferably not lower than 1/100 and not higher than
100.
[0018] According to the present invention, the content of the
lactoferrin and/or lactoferrin hydrolysate based on the whole
nutritional composition is preferably 0.001 to 10.0 mass %.
[0019] According to the present invention, the content of the human
milk oligosaccharide based on the entire nutritional composition is
preferably 0.001 to 10.0 mass %.
[0020] According to the present invention, the human milk
oligosaccharide preferably contains one or more of
2'-fucosyllactose, 3-fucosyllactose, 3'-sialyllactose, and
6'-sialyllactose.
[0021] The nutritional composition of the present invention can be
preferably used for preventing infection, and can be more
preferably used for preventing gastrointestinal tract
infection.
[0022] The nutritional composition of the present invention is
preferably in the form of a powdered formula or a liquid
formula.
[0023] The nutritional composition of the present invention is
preferably administered to infants, young children, or elderly
persons.
BRIEF DESCRIPTION OF THE DRAWINGS
[0024] FIG. 1 A graph showing infection-suppressing ratio for
rotavirus in cells to which lactoferrin and/or 2'-fucosyllactose
was added.
[0025] FIG. 2 A graph showing infection-suppressing ratio for
rotavirus in cells to which lactoferrin and/or 3-fucosyllactose was
added.
[0026] FIG. 3 A graph showing infection-suppressing ratio for
rotavirus in cells to which lactoferrin and/or 3-fucosyllactose was
added.
[0027] FIG. 4 A graph showing infection-suppressing ratio for
rotavirus in cells to which lactoferrin and/or 3'-sialyllactose was
added.
[0028] FIG. 5 A graph showing infection-suppressing ratio for
rotavirus in cells to which lactoferrin and/or 6'-sialyllactose was
added.
[0029] FIG. 6 A graph showing infection-suppressing ratio for
rotavirus in cells to which lactoferrin and/or 6'-sialyllactose was
added.
[0030] FIG. 7 A graph showing infection-suppressing ratio for
rotavirus in cells to which lactoferrin and/or human milk
oligosaccharides (mixture) were added.
EFFECT OF THE INVENTION
[0031] According to the present invention, a nutritional
composition suitable for use in prevention of infection is
provided. This nutritional composition can be contained in foods,
drinks, and drugs as an additive or the like. Further, use of the
nutritional composition of the present invention itself as a food
or drink is expected to be useful for health maintenance of
especially infants, young children, and elderly persons.
MODES FOR CARRYING OUT THE INVENTION
[0032] Hereafter, the present invention will be explained in
detail. However, the present invention is not limited to the
following embodiments but can be freely changed within the scope of
the present invention.
[0033] The nutritional composition of the present invention
contains lactoferrin and/or a lactoferrin hydrolysate, and a human
milk oligosaccharide.
[0034] Lactoferrin is an iron-binding glycoprotein contained in
milk, tears, saliva, blood, and so forth of mammals such as sheep,
goat, pig, mouse, water buffalo, camel, yak, horse, donkey, llama,
cow, and human.
[0035] Although lactoferrin used for the present invention may
originate in any mammal, and is not particularly limited, for
example, lactoferrin derived from the milk of a cow, human, or the
like is preferred in view of content and availability. The milk may
be any of colostrum, transitional milk, nature milk, and late
lactation milk.
[0036] Lactoferrin used for the present invention may be
lactoferrin separated from skim milk, whey, and so forth, which are
processed products of the milk, in a conventional manner (for
example, ion chromatography etc.), recombinant lactoferrin produced
by a genetically manipulated microorganism, animal cell, transgenic
animal, or the like, synthetic lactoferrin, or a mixture of these.
Lactoferrin may be a deglycosylated or glycosylated lactoferrin. As
such lactoferrin, marketed lactoferrin produced on an industrial
scale (produced by, for example, Morinaga Milk Industry Co., Ltd.)
can be used.
[0037] The metal content of lactoferrin used for the present
invention is not particularly limited, and any one of
apolactoferrin obtained by deferrization of lactoferrin with
hydrochloric acid, citric acid, or the like; metal-saturated
lactoferrin in a saturation degree of 100% or more obtained by
chelating the apolactoferrin with a metal such as iron, copper,
zinc, and manganese; and partially metal-saturated lactoferrin in
which metal binds at a saturation degree lower than 100%, or a
mixture consisting of two or more of these can be used.
[0038] Hereafter, an example of a method for preparing lactoferrin
(separation and purification of lactoferrin from a raw material
such as milk) used for the present invention will be explained
below. However, the method is not limited to this method.
[0039] A milk raw material derived from cow is loaded on a cation
exchange column, the eluate is collected, and this eluate is
repeatedly loaded on the column as required. Deionized water is
loaded on this column, and an aqueous sodium chloride solution is
further loaded to obtain an eluate containing basic proteins having
been adsorbed onto the cation exchange column. The proteins are
collected from this eluate by ammonium sulfate precipitation and
washed as required. The collected precipitates are dissolved in
deionized water, and this solution is filtered through an
ultrafiltration membrane. The residue can be further subjected to
desalting and lyophilization to obtain powdered lactoferrin.
[0040] More precisely, the column is filled with an ion exchanger,
hydrochloric acid is loaded, and the ion exchanger is washed with
water and thereby equilibrated. Then, skim milk of pH 6.9 cooled to
4.degree. C. is loaded onto the column, and the eluate is
collected, and similarly loaded on the column again. Subsequently,
deionized water is loaded on the column, and an aqueous sodium
chloride solution is further loaded to obtain an eluate containing
basic proteins having been adsorbed on the cation exchange column.
Ammonium sulfate at a saturation degree of 80% is added to this
eluate to precipitate the proteins, and the precipitates are
collected by centrifugation. The collected precipitates are washed
with an 80% saturated ammonium sulfate solution, and deionized
water is added to dissolve the precipitates; the obtained solution
is desalted by using an ultrafiltration membrane module, and the
residue is lyophilized to obtain powdered bovine lactoferrin.
Bovine lactoferrin having a purity of 95 mass % or higher can be
obtained as described above.
[0041] Lactoferrin genes naturally contain a genetic mutation such
as a substitution, deletion, insertion and inversion of one or more
nucleotides at one or more positions depending on the difference of
species, genus, individual, etc., and amino acid sequence of a
protein encoded by a gene containing such mutations may also
contain mutations. Lactoferrin used for the present invention may
be lactoferrin containing such a mutation, so long as the effect of
the present invention is not degraded.
[0042] Lactoferrin used for the present invention may also be a
processed product of lactoferrin obtained by heat treatment, acid
treatment, or alkaline treatment of lactoferrin, so long as the
effect of the present invention is not degraded.
[0043] The lactoferrin hydrolysate referred to in the present
invention is such lactoferrin as mentioned above subjected to a
hydrolysis treatment.
[0044] Examples of the hydrolysis treatment include, for example,
the hydrolysis treatment by the method described in Japanese Patent
Laid-open (KOKAI) No. 2012-235768.
[0045] Specifically, before a lactoferrin solution is subjected to
an enzymatic reaction treatment, the solution is adjusted to pH 2
to 4, preferably pH 2.5 to 3.5, particularly preferably pH 3, with
an acid such as hydrochloric acid, citric acid, and acetic
acid.
[0046] An enzyme composition containing chymosin is added in a
desired amount to the lactoferrin solution of which pH has been
adjusted, and then the temperature is maintained for the enzymatic
reaction at 35 to 55.degree. C., preferably 40 to 50.degree. C.,
more preferably 42 to 48.degree. C., with stirring to hydrolyze
lactoferrin for 6 to 24 hours, preferably 12 to 18 hours.
[0047] Subsequently, the temperature of the reaction solution is
elevated to, for example, 80.degree. C., and maintained at that
temperature for 10 minutes to carry out heat inactivation of the
enzyme. An alkali solution such as sodium hydroxide solution is
preferably further added to adjust the solution to pH 5 to 7, for
example, pH 6.
[0048] Although the reaction solution (lactoferrin hydrolysate) of
which pH has been adjusted may be used as solution, it is
preferably powdered by lyophilization, or the like. As the
lactoferrin hydrolysate, a fractionation product of the reaction
solution obtained by chromatography, ultrafiltration, or the like
can also be used.
[0049] In the nutritional composition of the present invention,
content of lactoferrin and/or lactoferrin hydrolysate based on the
entire nutritional composition is preferably 0.001 to 5.0 mass %,
more preferably 0.005 to 1.0 mass %, more preferably 0.01 to 0.5
mass %, particularly preferably 0.01 to 0.2 mass %.
[0050] These ranges may usually be the ranges of the content in the
nutritional composition that is to be distributed as a commercial
product.
[0051] The human milk oligosaccharide used for the present
invention is not particularly limited so long as it is an
oligosaccharide usually contained in human milk, but preferred
examples include neutral human milk oligosaccharides such as
2'-fucosyllactose, 3-fucosyllactose, lactodifucotetraose,
2',3-difucosyllactose, lacto-N-triose II, lacto-N-tetraose,
lacto-N-neotetraose, lacto-N-fucopentaose I,
lacto-N-neofucopentaose, lacto-N-fucopentaose II,
lacto-N-fucopentaose III, lacto-N-fucopentaose V,
lacto-N-neofucopentaose V, lacto-N-difucohexaose I,
lacto-N-difucohexaose II, 6'-galactosyllactose,
3'-galactosyllactose, lacto-N-hexaose, and lacto-N-neohexaose, and
acidic human milk oligosaccharides such as 3'-sialyllactose,
6'-sialyllactose, 3-fucosyl-3'-sialyllactose, and
disialyl-lacto-N-tetraose. Among these, 2'-fucosyllactose,
3-fucosyllactose, 3'-sialyllactose, and 6'-sialyllactose are
preferred, and 2'-fucosyllactose is particularly preferred.
[0052] The human milk oligosaccharide used for the present
invention may be a purified oligosaccharide or may be a mixture so
long as the effect of the present invention is not degraded. And
the human milk oligosaccharide used for the present invention may
contain one or more of the human milk oligosaccharides mentioned
above.
[0053] In the nutritional composition of the present invention, the
content of the human milk oligosaccharide based on the entire
nutritional composition is preferably 0.001 to 10.0 mass %, more
preferably 0.005 to 5.0 mass %, further preferably 0.005 to 2.0
mass %, particularly preferably 0.01 to 2.0 mass %.
[0054] In the composition of the present invention, mass ratio of
the human milk oligosaccharide to lactoferrin and/or lactoferrin
hydrolysate is preferably not lower than 1/100 and not higher than
100, more preferably not lower than 1/10 and not higher than 100,
further preferably not lower than 1/5 and not higher than 50.
[0055] These ranges may usually be the ranges of the content in the
nutritional composition that is to be distributed as a commercial
product.
[0056] The contents at the time of ingestion (administration) of
the nutritional composition of the present invention may be within
the aforementioned ranges, or the composition may be, for example,
diluted, as required. For example, the content of lactoferrin
and/or lactoferrin hydrolysate based on the entire composition at
the time of ingestion (administration) can be 0.0001 to 10.0 mass
%. Further, the content of the human milk oligosaccharide based on
the entire composition at the time of ingestion (administration)
can be 0.0001 to 10.0 mass %. The mass ratio of the human milk
oligosaccharide to lactoferrin and/or lactoferrin hydrolysate at
the time of ingestion (administration) can be not lower than 1/100
and not higher than 100.
[0057] The nutritional composition of the present invention may
further contain a probiotic.
[0058] As the probiotic used for the present invention,
Lactobacillus bacteria, Streptococcus bacteria, and Bifidobacterium
bacteria, which are resident lactic acid bacteria in the human
alimentary canal, can be mainly used. In particular, lactic acid
bacteria of the genus Lactobacillus are most frequently used.
Especially, Lactobacillus rhamnosus GG strain (Japanese Patent
Laid-open (KOKAI) 61-280433), Lactobacillus casei Shirota strain
(marketed by Yakult Honsha Co., Ltd.), Lactobacillus johnsonii La1
strain (Japanese Patent Laid-open (KOKAI) No. 6-315373),
Lactobacillus plantarum 299 strain (DSM6595) (Japanese Patent
Laid-open (KOHYO) No. 6-501624), Lactobacillus plantarum 299v
strain (DSM9843) (Japanese Patent Laid-open (KOHYO) No. 11-502703),
Lactobacillus salivarius UCC 1 strain (NCIMB 40830), Lactobacillus
salivarius UCC 118 strain (NCIMB 40829, WO98/35014), and so forth
can used as strains suitable for probiotic products.
[0059] The Lactobacillus bacteria are not particularly limited, and
examples include Lacticaseibacillus casei, Lacticaseibacillus
paracasei, Lacticaseibacillus rhamnosus, Lactiplantibacillus
plantarum, Levilactobacillus brevis, Ligilactobacillus salivarius,
Limosilactobacillus fermentum, Limosilactobacillus reuteri.
[0060] In the present invention, Bifidobacterium bacteria are still
more preferably used as the probiotic.
[0061] The Bifidobacterium bacteria are not particularly limited,
and examples include Bifidobacterium breve, Bifidobacterium
infantis (later reclassified as Bifidobacterium longum subsp.
infantis), Bifidobacterium longum, Bifidobacterium bifidum,
Bifidobacterium adolescentis, Bifidobacterium catenulatum,
Bifidobacterium pseudocatenulatum, Bifidobacterium animalis,
Bifidobacterium lactis, and Bifidobacterium pseudolongum. Among
these, one or more of Bifidobacterium breve, Bifidobacterium
infantis, and Bifidobacterium longum are preferred.
[0062] In the present invention, the probiotic, more preferably
Bifidobacterium bacteria, may consist of live cells or dead cells.
When live cells are used, 1 mL of the nutritional composition
preferably contains 1.0.times.10.sup.6 cfu or more, more preferably
1.0.times.10.sup.7 cfu or more, further preferably
2.0.times.10.sup.7 cfu or more, of live cells. When live cells are
used, dead cells may also be present.
[0063] The unit "cfu" means "colony-forming unit." In this
description, the values of this unit may be, for example, those
obtained in a culture at 38.degree. C. on a solid medium containing
10 mass % of reduced skim milk powder.
[0064] Preferred examples of the Bifidobacterium bacteria to be
present in the nutritional composition of the present invention are
one or more of Bifidobacterium breve, Bifidobacterium infantis, and
Bifidobacterium longum.
[0065] More specifically, examples of Bifidobacterium breve include
Bifidobacterium breve M-16V. Bifidobacterium breve M-16V was
deposited on Jan. 26, 2018 at the independent administrative
agency, National Institute of Technology and Evaluation, NITE
Patent Microorganisms Depositary (NPMD, #122, 2-5-8, Kazusa
Kamatari, Kisarazu-shi, Chiba-ken, 292-0818, Japan) as an
international deposit under the provisions of the Budapest Treaty,
and given the accession number of NITE BP-02622. Commercially
available products, for example, "Bifidobacterium breve M-16V"
produced by Morinaga Milk Industry Co., Ltd., may be used.
[0066] More specifically, examples of Bifidobacterium breve also
include Bifidobacterium breve MCC1274. Bifidobacterium breve
MCC1274 was deposited on Aug. 25, 2009 at the independent
administrative agency, National Institute of Advanced Industrial
Science and Technology, International Patent Organism Depositary
(currently, the independent administrative agency, National
Institute of Technology and Evaluation, International Patent
Organism Depositary (IPOD, #120, 2-5-8, Kazusa Kamatari,
Kisarazu-shi, Chiba-ken, 292-0818, Japan) as an international
deposit under the provisions of the Budapest Treaty, and given the
accession number of FERM BP-11175.
[0067] More specifically, examples of Bifidobacterium infantis also
include Bifidobacterium infantis M63. Bifidobacterium infantis M63
was deposited on Jan. 26, 2018 at NPMD as an international deposit
under the provisions of the Budapest Treaty, and given the
accession number of NITE BP-02623.
[0068] More specifically, as Bifidobacterium longum,
Bifidobacterium longum NITE BP-02621 (synonym: BB536 or
Bifidobacterium longum subsp. longum ATCC BAA-999) can be used.
Bifidobacterium longum BB536 was deposited at NPMD on Jan. 26, 2018
as an international deposit under the provisions of the Budapest
Treaty, and given the accession number of NITE BP-02621. The same
bacterial strain, Bifidobacterium longum subsp. longum ATCC BAA-999
(ATCC BAA-999), can be obtained from American Type Culture
Collection (ATCC, 10801 University Boulevard, Manassas, Va. 20110,
United States of America) as ATCC BAA-999 (see, for example,
Japanese Patent Laid-open (KOKAI) No. 2012-223134 etc.).
[0069] The bacteria identified by the names of the bacteria
exemplified above are not limited to the strains deposited or
registered at the depositaries mentioned above themselves
(henceforth referred to as "deposited strains" for convenience of
explanation), and may be strains substantially equivalent to the
deposited strains (also referred to as "derived strains" or
"derivative strains"). The "strains substantially equivalent to the
aforementioned deposited strains" refer to strains that belong to
the same species as those of the aforementioned deposited strains,
provide an intestinal bacterial flora-improving effect, preferably
have a nucleotide sequence of the 16SrRNA gene showing an identity
of 99.86% or higher, more preferably 99.93% or higher, further
preferably 100%, to the nucleotide sequence of 16SrRNA genes of the
aforementioned deposited strains, and preferably have the same
bacteriological characteristics as those of the aforementioned
deposited strains, respectively. The strains substantially
equivalent to the aforementioned deposited strains may be, for
example, derivative strains derived from the respective deposited
strains as a parent strain. Examples of the derivative strains
include a strain bred from any of the deposited strains, and a
strain naturally produced from any of the deposited strains.
Examples of the breeding method include modification based on a
genetic engineering technique and modification caused by using a
mutation treatment. Examples of the mutation treatment include
irradiation of X-rays, irradiation of ultraviolet rays, and
treatment with a mutation agent such as
N-methyl-N'-nitro-N-nitrosoguanidine, ethyl methanesulfonate, and
methyl methanesulfonate. Examples of the strain naturally produced
from any of the deposited strains include a strain naturally
produced at the time of use of any of the deposited strains.
Examples of such a strain include a mutant strain naturally
produced in culture (for example, subculture) of any of the
deposited strains. The derivative strains may be constructed by one
type of modification, or may be constructed by two more types of
modifications.
[0070] As cells of Bifidobacterium bacteria that may be present in
the nutritional composition of the present invention, commercial
products may be used, or those obtained by an appropriate
production method may be used.
[0071] Cells of Bifidobacterium bacteria that may be present in the
nutritional composition of the present invention can also be easily
obtained by culturing any of the aforementioned Bifidobacterium
bacteria. The culture method is not particularly limited so long as
Bifidobacterium bacteria can be proliferated. As the culture
method, for example, methods usually used for culturing
Bifidobacterium bacteria can be used as they are, or with
appropriate modifications. The culture temperature may be, for
example, 25 to 50.degree. C., preferably 35 to 42.degree. C. The
culture can be preferably carried out under anaerobic conditions,
and for example, it can be carried out by supplying an anaerobic
gas such as carbon dioxide gas. The culture can also be carried out
under microaerobic conditions such as those obtainable by
stationary liquid culture or the like. The culture can be carried
out, for example, until Bifidobacterium bacteria proliferate to a
desired level.
[0072] The medium used for the culture is not particularly limited
so long as Bifidobacterium bacteria can be proliferated. As the
medium, for example, media usually used for culturing
Bifidobacterium bacteria can be used as they are or with
appropriate modifications. That is, as the carbon source, for
example, saccharides such as galactose, glucose, fructose, mannose,
cellobiose, maltose, lactose, sucrose, trehalose, starch, starch
hydrolysate, and blackstrap molasses can be used depending on
assimilation property of the bacteria. As the nitrogen source, for
example, ammonia, ammonium salts such as ammonium sulfate, ammonium
chloride, and ammonium nitrate, and nitrates can be used. As
inorganic salts, for example, sodium chloride, potassium chloride,
potassium phosphate, magnesium sulfate, calcium chloride, calcium
nitrate, manganese chloride, ferrous sulfate, and so forth can be
used. Organic components such as peptone, soybean flour, defatted
soybean meal, meat extract, and yeast extract may also be used.
Specifically, examples of media usually used for culturing
Bifidobacterium bacteria include the reinforced clostridial medium,
MRS medium (de Man, Rogosa, and Sharpe medium), mMRS medium
(modified MRS medium), TOSP medium (TOS propionate medium), and
TOSP Mup medium (TOS propionate mupirocin medium).
[0073] As Bifidobacterium bacteria that may be contained in the
nutritional composition of the present invention, cells of the
bacteria or a fraction containing the cells can be used without any
particular limitation. That is, as Bifidobacterium bacteria, for
example, a culture obtained by culturing the bacteria can be used
as it is, such culture may be used after dilution or concentration,
or cells collected from such culture may be used. So long as the
intestinal bacterial flora-improving effect is not degraded, the
culture product may be subjected to any of various additional
treatments such as heating and lyophilization after the culture.
Additional treatments that allow higher survivability of the cells
are more preferred. That is, specific examples of Bifidobacterium
bacteria that may be present in the nutritional composition of the
present invention include culture of Bifidobacterium bacteria,
cells collected from culture of Bifidobacterium bacteria, and
processed products of these, and examples of the processed products
include products obtained by dilution, concentration, drying, or
the like. The cells usually preferably include live cells. The
cells may consist of, for example, live cells or a mixture of live
cells and dead cells.
[0074] The nutritional composition of the present invention may
further contain a prebiotic.
[0075] The term "prebiotic" may indicate an indigestible food
component that allows selective proliferation of useful bacteria in
the intestinal tract. Examples of such useful bacteria include
Bifidobacterium bacteria. That is, as the prebiotic, an ingredient
that selectively proliferates Bifidobacterium bacteria is
preferred. As the prebiotic, one kind of ingredient may be used, or
a combination of two or more kinds of ingredients may be used.
[0076] The prebiotic is not particularly limited by, specifically,
a probiotic concomitantly used, so long as the desired effect can
be obtained. Examples of the prebiotic include oligosaccharides,
dietary fibers, and gluconic acid. Examples of the oligosaccharides
include galactooligosaccharides, fructooligosaccharides,
xylooligosaccharide, isomaltooligosaccharides, raffinose,
lactulose, lactosucrose, soybean oligosaccharides, and coffee
oligosaccharides.
[0077] The nutritional composition of the present invention can be
used for treating an infectious disease or pathological condition,
wherein the nutritional composition can prevent infection or
stimulate immunity. It can be used especially as a nutritional
composition for preventing viral infection.
[0078] The expression "for prevention of viral infection" used for
the present invention usually indicates a use for making infants,
young children and elderly persons with low resistance who are
easily infected with viruses, exhibit an effect of preventing
infection with virus, or protecting them from the infection.
Examples of viruses for which such a protection effect for the
infection thereof can be exhibited include rotaviruses and most
viruses of the family Reoviridae to which rotaviruses belong,
influenza viruses, noroviruses, cytomegaloviruses, RS viruses,
polioviruses, and HIV.
[0079] The expression "action for preventing infection" used in
this description refers to an action of the nutritional composition
of the present invention of preventing viral infection or the
action of mitigating the pathological symptoms in infected
patients, through ingestion of the composition before and during
the epidemic period of viral infection.
[0080] The nutritional composition of the present invention can be
preferably used for the prevention of viral infection in the
gastrointestinal tract. Specific examples of virus that can infect
gastrointestinal tract include rotaviruses and noroviruses. It is
considered that the action mechanism of the prevention of infection
by the nutritional composition of the present invention is based on
suppression of adhesion of the viruses with cells, specifically,
cells of gastrointestinal tract, more specifically, intestinal
epithelial cells. It is also considered that the action mechanism
of the prevention of infection (including reduction of severity) by
the nutritional composition of the present invention is also based
on suppression of gene replication of viruses in vivo after the
viral infection.
[0081] By adding an effective amount of the nutritional composition
as a nutritional composition for childcare of infants, young
children with immature defensive ability, or a food or nutritional
composition for elderly persons with reduced defensive ability
against infection, a nutritional composition for preventing
infection can be prepared.
[0082] Another aspect of the present invention is the use of
lactoferrin and/or a lactoferrin hydrolysate, and a human milk
oligosaccharide in the manufacture of a nutritional composition for
preventing infection.
[0083] Still another aspect of the present invention is use of a
nutritional composition containing lactoferrin and/or a lactoferrin
hydrolysate, and a human milk oligosaccharide for preventing
infection.
[0084] Still another aspect of the present invention is a
nutritional composition containing lactoferrin and/or a lactoferrin
hydrolysate, and a human milk oligosaccharide, which is used for
preventing infection.
[0085] Still another aspect of the present invention is a method
for preventing infection, which includes administering lactoferrin
and/or a lactoferrin hydrolysate, and a human milk oligosaccharide
to an animal or human. Although the object of the administration is
not particularly limited, it is usually a human.
[0086] The ingestion (administration) period of the nutritional
composition of the present invention is not particularly limited
and can be appropriately chosen according to the condition of the
object of the administration.
[0087] The ingestion (administration) amount of the nutritional
composition of the present invention is appropriately chosen
according to age, sex, pathological condition, other conditions,
and so forth of a subject of the ingestion (object of
administration).
[0088] The nutritional composition of the present invention is
preferably prepared in the form of food or drink.
[0089] Forms and properties of the food and drink are not
particularly limited so long as the effect of the present invention
is not degraded, and they can be prepared by usual methods using
raw materials usually used for foods and drinks.
[0090] The nutritional composition in the form of an additive to be
added to a food or drink also falls within the scope of the
composition of the present invention. Examples of this embodiment
include, for example, additives to be added to collected mother's
milk or formula milk, and ingestion of the milk containing the
additive by neonates or infants is supposed.
[0091] Although food or drink is usually orally ingested, the
ingestion route is not limited to this route, and for example, it
may be ingested via a paranasal route, or ingested by using a
gastric fistula or cecal fistula. For example, it is supposed that
formula milk for neonates or infants, which is the composition of
the present invention mentioned later, or mother's milk to which
the composition of the present invention is added is ingested by
neonates or infants through a paranasal stomach feeding tube, or
the like.
[0092] Forms of the food and drink are not particularly limited,
and they may be in the form of liquid, paste, gellated solid,
powder, or the like. Examples include, for example, tablet
confectioneries, and liquid diets (nutrition diets for tube
feeding), as well as flour products such as bread, macaroni,
spaghetti, noodles, cake mix, fry powder and bread crumbs;
ready-to-eat foods such as instant noodles, cup noodles, retort and
cooked foods, canned cooking, foods for microwave heating, instant
soup and stew, instant miso soup and Japanese clear soup, canned
soup, freeze-dried foods, and other ready-to-eat foods; processed
agricultural products such as canned agricultural products, canned
fruits, jams and marmalades, pickles, cooked beans, dry
agricultural products, and cereals (processed grain products);
processed marine products such as canned marine products, fish hams
and sausages, seafood paste products, marine dainties, and
tsukudani (marine products boiled in soy sauce); processed
livestock products such as canned livestock products and pastes,
and livestock meat hams and sausages; milks and dairy products such
as processed milk, milk drinks, yoghurts, lactic acid drinks,
cheese, ice creams, creams, and other dairy products; oils and fats
such as butters, margarines, and vegetable oils; basic seasonings
such as soy sauce, miso, sauces, processed tomato seasoning, mirin,
and vinegars; complex seasonings and foods such as cooking mix,
curry powder or roux, sauces for dipping, dressings, noodle soups,
spices, and other complex seasonings; frozen foods such as frozen
food materials, semi-cooked frozen foods, and cooked frozen foods;
confectioneries such as caramels, candies, chewing gums,
chocolates, cookies, biscuits, cakes, pies, snacks, crackers,
Japanese sweets, rice confectioneries, bean confectioneries,
dessert pastries, jellies, and other confectioneries; beverages
such as carbonated drinks, natural fruit juices, fruit juice
drinks, fruit juice soft drinks, fruit pulp drinks, fruit drinks
with fruit pulp, vegetable based drinks, soy milk, soy milk drinks,
coffee drinks, tea drinks, powdered drinks, concentrated drinks,
sports drinks, energy drinks, alcoholic drinks, and other
beverages; other commercial foods such as baby foods, rice
seasonings, and seaweed seasonings for boiled rice soaked with tea;
nutritional compositions such as supplements and formula milk
(including powdered formula and liquid formula); nutritional
compositions for infants such as supplements for infants and infant
formula(including powdered formula and liquid formula); enteral
nutrition; functional foods (foods for specified health uses, foods
with nutrient function claims), and so forth.
[0093] Among these, a nutritional composition for infants and
children is preferred.
[0094] In Japan, the term nutritional composition for infants and
children refers to a nutritional composition for 0 to 12 months old
infants (also referred to as infant formula), follow-up milk for 6
to 9 months old infants and children (up to 6 years old) (also
referred to as formula for infants and children), nutritional
composition for low birth weight infants of birth weight lower than
2500 g (low birth weight infants) (also referred to as formula for
premature or low birth weight infants), formulas for special
medical purpose used for treating infants with pathological
conditions such as cow's milk allergy and lactose intolerance, and
so forth. The nutritional composition can also be applied to foods
with health claims or foods for medical purpose.
[0095] Outside Japan, the term nutritional composition for infants
and children refers to a nutritional composition for 0 to 6 month
old infants (also referred to as infant formula), nutritional
composition for 6 to 12 month old infants and for 1 to 3 year old
children (also referred to as follow-up formula), nutritional
composition for 3 to 6 year old children (also referred to as
growing-up milk), nutritional composition for low birth weight
infants of birth weight lower than 2500 g (also referred to as
formula for premature or low birth weight infants), formulas for
special medical purpose used for treating infants with pathological
conditions such as cow's milk allergy and lactose intolerance, and
so forth. The nutritional composition can also be applied to foods
with health claims or foods for medical purpose.
[0096] Examples of the nutritional composition of the present
invention also include balanced nutritional foods such as formula
milk for pregnant and lactating mothers (formula milk containing
nutrients required for gestation period and lactation period in
good balance), formula milk for school-age children, formula milk
for adults, and liquid foods for elderly persons, nutritional
supplements, and foods for medical purpose such as
phosphorus-reduced powdered milk (food for special medical
purpose).
[0097] As an embodiment of food or drink, the nutritional
composition of the present invention may be prepared as a feed.
Examples of the feed include pet foods, feeds for livestock, feeds
for fish breeding, and so forth.
[0098] Forms of the feeds are not particularly limited, and the
feeds may be, for example, those containing cereals such as corn,
wheat, barley, rye, and milo; vegetable oil cakes such as soybean
oilcake, rapeseed oilcake, coconut oilcake, and linseed oilcake;
brans such as wheat bran, rice bran, and defatted rice bran;
manufacturing meals such as corn gluten meal and corn germ meal;
animal feeds such as fish meal, skim milk powder, whey, yellow
grease, and tallow; yeasts such as torula yeast and brewer's yeast;
mineral feeds such as tribasic calcium phosphate and calcium
carbonate; oils and fats; simple amino acids; saccharides, and so
forth.
[0099] When the nutritional composition of the present invention is
in the form of food or drink, it may be a food with health claims
or a food for special dietary uses. The food with health claims
system was provided not only for usual foods, but also for foods in
the form of tablet, capsule, or the like in consideration of the
domestic and foreign trends and the compatibility with the
conventional food for specified health uses system, and foods with
health claims are classified into three types, foods for specified
health uses, foods with function claims, and foods with nutrient
function claims. The foods for special dietary uses are foods for
special uses for those who cannot eat usual meals such as sick
persons, infants, young children, and elderly persons, and include
foods for special medical purpose (standard regulation type and
individual evaluation type), powdered formula for pregnant women
and lactating mothers, infant formula, and foods for dysphagia.
[0100] When the composition of the present invention is in the form
of food or drink (including feed), it can be provided and sold as a
food or drink with an indication (labeling) of use for preventing
infection.
[0101] The aforementioned term "indication" means all actions for
informing consumers the aforementioned use, and any indications
reminding or analogizing the aforementioned use fall within the
scope of the "indication" referred to in the present invention
regardless of purpose and content of the indication, objective
article or medium on which the indication is used, and so
forth.
[0102] The term "indication" is preferably made with an expression
that allows consumers to directly recognize the aforementioned use.
Specific examples include actions of assigning or delivering
commodities including the food or drink on which the aforementioned
use is indicated or such commodities having packages on which the
aforementioned use is indicated, as well as displaying or importing
such commodities, displaying or distributing advertisements, price
lists or business papers relating to such commodities on which the
aforementioned use is indicated, providing information including
those as contents that indicate the aforementioned use by an
electromagnetic method (Internet etc.) for the purpose of assigning
or delivering such commodities, and so forth.
[0103] The indication is preferably an indication approved by an
administration etc. (for example, an indication based on an
approval, which is qualified on the basis of any of various legal
systems provided by the government of Japan). Indications of such
contents on packages, containers, catalogues, pamphlets,
advertisement materials used at the point of sale, such as POPs,
and other papers are preferred.
[0104] Examples of the "indication" include, for example,
indications as health food, functional food, enteral nutritive
food, food for special dietary uses, food with health claims, food
for specified health uses, food with nutrient function claims, food
with function claims, quasi-drug, and so forth. Among these,
indications approved by the Consumer Affairs Agency, for example,
indications approved on the basis of the systems of food for
specified health uses, food with nutrient function claims, food
with function claims, and similar systems can be especially
mentioned. Specific examples include indications as food for
specified health uses, indications as food for specified health
uses with qualified health claims, indications of influence on body
structures and functions, indications of reduction of disease risk
claims, indications of functional claims based on scientific
evidences, and so forth, and more precisely, typical examples
include the indications as food for specified health uses
(especially indications of use for health) defined in the Cabinet
Office Ordinance concerning approval of indications of special
dietary uses provided by the Health Promotion Law, and others
(Japanese Cabinet Office Ordinance No. 57, Aug. 31, 2009), and
similar indications.
[0105] As such an indication, for example, wordings of "for
preventing infection", "measure against viral infection", "for
infants' health", and so forth can be indicated.
[0106] The composition of the present invention can also be in the
form of drug, and in this case, as for the administration route
thereof, it may be orally or parenterally administered, but it is
preferably orally administered. Examples of parenteral ingestion
(administration) include rectal administration, and so forth.
[0107] As for form of the drug, it can be appropriately prepared in
a desired dosage form depending on the administration method. For
example, in the case of oral administration, it can be prepared in
the form of solid preparation such as a powder, granule, tablet,
and capsule; liquid agent such as a solution, syrup, suspension,
and emulsion, or the like. In the case of parenteral
administration, it can be prepared in the form of a suppository,
ointment, injection, or the like.
[0108] When the drug is prepared, such ingredients as excipients,
pH adjustors, colorants and corrigents, which are usually used for
the preparation of drugs, can be used. Another pharmaceutical
ingredient, a prebiotic for Bifidobacterium bacteria that is known,
or one that is discovered in the future, or the like may also be
used together.
[0109] The drug can be prepared by a known method appropriately
chosen depending on the dosage form. The drug may also be prepared
by adding a carrier for pharmaceutical compositions.
[0110] Examples of excipients include, for example, saccharides and
derivatives thereof such as lactose, sucrose, glucose, mannitol,
and sorbitol; starch and derivatives thereof such as corn starch,
potato starch, pregelatinized starch, dextrin, and carboxymethyl
starch; cellulose and derivatives thereof such as crystalline
cellulose, hydroxypropylcellulose, hydroxypropylmethylcellulose,
carboxymethylcellulose and carboxymethylcellulose calcium; gum
arabic; dextran; pullulan; silicate and derivatives thereof such as
light silicic anhydride, synthetic aluminum silicate and magnesium
metasilicate aluminate; phosphate and derivatives thereof such as
calcium phosphate; carbonate and derivatives thereof such as
calcium carbonate; sulfate and derivatives thereof such as calcium
sulfate, and so forth.
[0111] Examples of binders include, in addition to the
aforementioned excipients, for example, gelatin;
polyvinylpyrrolidone; Macrogol, and so forth.
[0112] Examples of disintegrating agents include, in addition to
the aforementioned excipients, for example, chemically modified
starch or cellulose derivatives such as croscarmellose sodium, and
carboxymethylstarch sodium, crosslinked polyvinylpyrrolidone, and
so forth.
[0113] Examples of lubricants include, for example, talc; stearic
acid; stearic acid metal salts such as calcium stearate and
magnesium stearate; colloidal silica; waxes such as peegum and
spermaceti; boric acid; glycols; carboxylic acids such as fumaric
acid and adipic acid; carboxylic acid sodium salts such as sodium
benzoate; sulfates such as sodium sulfate; leucine; laurylsulfates
such as sodium laurylsulfate and magnesium laurylsulfate; silicic
acids such as silicic anhydride and silicic acid hydrate; starch
derivatives, and so forth.
[0114] Examples of stabilizers include, for example, p-oxybenzoic
acid esters such as methylparaben and propylparaben; alcohols such
as chlorobutanol, benzyl alcohol and phenylethyl alcohol;
benzalkonium chloride; acetic anhydride; sorbic acid, and so
forth.
[0115] Examples of corrigents include, for example, sweeteners,
acidulants, perfumes, and so forth.
[0116] Examples of carriers used for liquids for oral
administration include solvents such as water, and so forth.
EXAMPLES
[0117] Hereafter, the present invention will be explained more in
detail with reference to examples. However, the present invention
is not limited by these examples.
Example 1
[0118] Examination of antiviral action exhibited by lactoferrin,
human milk oligosaccharide, or both against simian rotavirus
[0119] The following experiment was performed to evaluate the
antiviral action exhibited by lactoferrin, human milk
oligosaccharide, or both against rotavirus.
[0120] The simian rotavirus SA11 strain (Nihon Bioresearch Inc.)
was inoculated to the MA104 cells (Sigma-Aldrich) of African green
monkey kidney maintained in a minimum essential medium
(Sigma-Aldrich, henceforth referred to simply as "medium"). The
simian rotavirus SA11 strain, for example, can be obtained from
American Type Culture Collection (ATCC) as ATCC VR-1565. To the
medium, lactoferrin (LF) and/or the human milk oligosaccharide(s)
shown below were added beforehand alone or in combination at the
following concentrations, respectively (n=3).
[0121] LF (produced by Morinaga Milk Industry Co., Ltd.): 0, 0.1 or
1.0 mg/mL
[0122] 2'-Fucosyllactose (2'-FL, Funakoshi Co., Ltd.): 0, 0.1, 0.5,
or 2.5 mg/mL
[0123] 3-Fucosyllactose (3-FL, Funakoshi Co., Ltd.): 0, 0.1, or 0.5
mg/mL
[0124] 3'-Sialyllactose (3'-SL, Funakoshi Co., Ltd.): 0, 0.1, or
0.5 mg/mL
[0125] 6'-Sialyllactose (6'-SL, Funakoshi Co., Ltd.): 0, 0.1, or
0.5 mg/mL
[0126] Human milk oligosaccharides (mixture) (HMO mix, mixture of
2'-FL (2.5 mg/mL), 3-FL (0.5 mg/mL), 3'-SL (0.25 mg/mL) and 6'-SL
(0.5 mg/mL))
[0127] The amount of 1.0 mg/mL corresponds to 0.1 mass % in mass
percentage.
[0128] Degree of infection was evaluated based on the number of
"plaques" formed when the cells were infected with the virus.
Specifically, the antiviral effects of lactoferrin and human milk
oligosaccharides were evaluated based on "plaque-suppressing ratio
(%)", i.e., the ratio of "plaque counts observed with lactoferrin
and/or human milk oligosaccharide(s) supplementation" to "plaque
counts observed without these supplementation". Plaque-suppressing
ratio was then regarded as "infection-suppressing ratio (%)".
[0129] The results are shown in FIGS. 1 to 7.
[0130] As shown in FIG. 1, the infection-suppressing ratio was 8.7%
when LF was used alone in an amount of 0.1 mg/mL, or 1.2% when
2'-FL was used alone in an amount of 0.1 mg/mL, but it was 15% when
0.1 mg/mL of LF and 0.1 mg/mL of 2'-FL were used in
combination.
[0131] As shown in FIG. 2, the infection-suppressing ratio was 8.7%
when LF was used alone in an amount of 0.1 mg/mL, or 6.6% when 3-FL
was used alone in an amount of 0.1 mg/mL, but it was 20% when 0.1
mg/mL of LF and 0.1 mg/mL of 3-FL were used in combination.
[0132] As shown in FIG. 3, the infection-suppressing ratio was 8.7%
when LF was used alone in an amount of 0.1 mg/mL, or 21% when 3-FL
was used alone in an amount of 0.5 mg/mL, but it was 32% when 0.1
mg/mL of LF and 0.5 mg/mL of 3-FL were used in combination.
[0133] As shown in FIG. 4, the infection-suppressing ratio was 8.7%
when LF was used alone in an amount of 0.1 mg/mL, or 33% when 3'-SL
was used alone in an amount of 0.5 mg/mL, but it was 41% when 0.1
mg/mL of LF and 0.5 mg/mL of 3'-SL were used in combination.
[0134] As shown in FIG. 5, the infection-suppressing ratio was 8.7%
when LF was used alone in an amount of 0.1 mg/mL, or 4.5% when
6'-SL was used alone in an amount of 0.1 mg/mL, but it was 14% when
0.1 mg/mL of LF and 0.1 mg/mL of 6'-SL were used in
combination.
[0135] As shown in FIG. 6, the infection-suppressing ratio was 8.7%
when LF was used alone in an amount of 0.1 mg/mL, or 23% when 6'-SL
was used alone in an amount of 0.5 mg/mL, but it was 30% when 0.1
mg/mL of LF and 0.5 mg/mL of 6'-SL were used in combination.
[0136] As shown in FIG. 7, the infection-suppressing ratio was 8.7%
when LF was used alone in an amount of 0.1 mg/mL, or 37% when the
HMO mix was used alone, but it was 46% when 0.1 mg/mL of LF and the
HMO mix were used in combination. Further, the
infection-suppressing ratio was 39% when LF was used alone in an
amount of 1.0 mg/mL, but it was 60% when 1.0 mg/mL of LF and the
HMO mix were used in combination.
[0137] As described above, when lactoferrin and human milk
oligosaccharides were added in combination, higher
plaque-suppressing ratios (infection-suppressing ratio) were
observed compared with those observed with adding lactoferrin alone
or human milk oligosaccharides alone. Therefore, it was recognized
that the combination of lactoferrin and human milk oligosaccharides
can exhibit synergistic or additive antiviral action against
rotavirus.
Preparation Example 1
[0138] A solution obtained by simultaneously dissolving lactoferrin
(produced by Morinaga Milk Industry Co., Ltd.) and human milk
oligosaccharides in water is lyophilized to obtain lyophilized
powder of a mixture of lactoferrin and human milk oligosaccharides.
The powder and whey protein concentrate (WPC, produced by Morinaga
Milk Industry Co., Ltd.) are uniformly mixed to obtain a
composition. This composition (20 g) is dissolved in water (200 g)
to obtain a nutritional composition for preventing viral
infection.
[0139] By administering the nutritional composition produced as
described above to a target person, viral infection of the target
person can be prevented.
Preparation Example 2
[0140] A solution obtained by simultaneously dissolving lactoferrin
(produced by Morinaga Milk Industry Co., Ltd.) and human milk
oligosaccharides in water is lyophilized to obtain a lyophilized
powder of a mixture of lactoferrin and human milk oligosaccharides.
The powder and dry powder of milk protein concentrate (MPC480,
produced by Fonterra Limited, protein content 80%, casein:whey
proteins=about 8:2)) are uniformly mixed to obtain a composition.
This composition (20 g) is dissolved in water (200 g) to obtain a
nutritional composition for preventing viral infection.
[0141] By administering the nutritional composition produced as
described above to a target person, viral infection of the target
person can be prevented.
Preparation Example 3
[0142] A solution obtained by simultaneously dissolving lactoferrin
(produced by Morinaga Milk Industry Co., Ltd.) and human milk
oligosaccharides in water is lyophilized to obtain lyophilized
powder of a mixture of lactoferrin and human milk oligosaccharides.
Subsequently, the powder and crystalline cellulose are put into a
stirring granulation machine, and mixed. Then, purified water is
added, the mixture is granulated, and the granulation product is
dried to obtain a granulated nutritional composition containing
lactoferrin and human milk oligosaccharides, as well as the
excipient.
[0143] By administering the nutritional composition produced as
described above to a target person, viral infection of the target
person can be prevented.
Preparation Example 4
[0144] A method for producing fermented milk to which lactoferrin
and human milk oligosaccharides are added will be explained
below.
[0145] First, a milk raw material, lactoferrin (produced by
Morinaga Milk Industry Co., Ltd.), and human milk oligosaccharides,
as well as water, other ingredients, etc. as required are mixed,
preferably homogenized, and sterilized by heating. The
homogenization and heat sterilization treatments can be performed
by conventional methods. A lactic starter is added (inoculated) to
the heat-pasteurized sterilized modified milk liquid, and the
modified milk liquid is maintained at a predetermined fermentation
temperature and thereby fermented to obtain a fermentation product.
Curd is formed by the fermentation.
[0146] As the lactic starter, for example, lactic acid bacteria
usually used for yogurt production such as Lactobacillus
bulgaricus, Lactococcus lactis, and Streptococcus thermophilus can
be used. If pH reaches a target value, the formed curd is crushed
by stirring, and cooled to 10.degree. C. or lower to obtain
fermented milk. By cooling to 10.degree. C. or lower, the activity
of the lactic acid bacterium can be reduced, and generation of the
acid can be suppressed.
[0147] Concentrated fermented milk can further be obtained by the
following method. The concentration step can be performed by
appropriately using a known concentration method. For example, a
centrifugation method or membrane separation method can be used. By
the centrifugation method, whey in the object of the concentration
(fermentation product containing lactoferrin and human milk
oligosaccharides) is removed, and concentrated fermented milk
containing lactoferrin and human milk oligosaccharides can be
obtained with a higher solid concentration.
[0148] A fermented milk containing lactoferrin and human milk
oligosaccharides according to the present invention can be produced
as described above.
[0149] By administering the fermented milk produced as described
above to a target person, viral infection of the target person can
be prevented.
Preparation Example 5
[0150] A method for producing a nutritional composition to which
lactoferrin and human milk oligosaccharides are added will be
explained below.
[0151] Desalted cow's milk whey protein powder (produced by Milei
GmbH, 10 kg), milk casein powder (produced by Fonterra Limited, 6
kg), lactose (produced by Milei GmbH, 48 kg), mineral mixture
(produced by Tomita Pharmaceutical Co., Ltd., 920 g), vitamin
mixture (produced by Tanabe Seiyaku Co., Ltd., 32 g), lactoferrin
(produced by Morinaga Milk Industry Co., Ltd., 17 to 690 g), and
human milk oligosaccharides (68 to 1698 g) are dissolved in warm
water (300 kg), the solution is heated at 90.degree. C. for 10
minutes to further dissolve the ingredients, modified fat (produced
by TAIYO YUSHI CORP., 28 kg) is added to the solution, and the
mixture is homogenized. Then, the homogenized mixture is subjected
to sterilization and concentration steps, and spray-dried to
prepare a nutritional composition containing lactoferrin and human
milk oligosaccharides (about 95 kg). If the obtained nutritional
composition is dissolved in water to prepare liquid formula having
a total solid concentration of 14% (w/v), which is the standard
solid concentration of formula milk, the lactoferrin concentration
and human milk oligosaccharide concentration in the formula milk
liquid will be 0.025 to 1 mg/mL and 0.1 to 5 mg/mL, respectively.
By administering the nutritional composition produced as described
above to an infant or young child, viral infection of the infant or
young children can be prevented.
Preparation Example 6
[0152] Lactoferrin (produced by Morinaga Milk Industry Co., Ltd.)
is dissolved in purified water at a concentration of 5 mass %, and
pH of the solution is adjusted to 3 with a hydrochloric acid
solution to prepare a lactoferrin solution. The prepared
lactoferrin solution is warmed to 45.degree. C., then calf rennet
(produced by RENCO Inc., containing 92 mass % of chymosin and 8
mass % of pepsin) is added in an amount of 3% based on the mass of
lactoferrin, and the reaction is allowed to occur for 24 hours with
stirring to cause hydrolysis.
[0153] After completion of the hydrolysis, the reaction mixture is
warmed to 80.degree. C., and maintained at the same temperature for
10 minutes to inactivate the enzymes.
[0154] The reaction mixture is cooled on ice, then a sodium
hydroxide solution is added to the reaction mixture to adjust pH of
the mixture to 6, and then the reaction mixture is lyophilized to
produce lactoferrin hydrolysate powder.
[0155] A solution obtained by simultaneously dissolving the
obtained lactoferrin hydrolysate powder and human milk
oligosaccharides in water is lyophilized to obtain lyophilized
powder of a mixture of the lactoferrin hydrolysate and human milk
oligosaccharides. The powder and whey protein concentrate (WPC,
produced by Morinaga Milk Industry Co., Ltd.) are uniformly mixed
to obtain a composition. This composition (20 g) is dissolved in
water (200 g) to obtain a nutritional composition for preventing
viral infection.
[0156] By administering the nutritional composition produced as
described above to a target person, viral infection of the target
person can be prevented.
Preparation Example 7
[0157] A method similar to the method for producing lactoferrin
hydrolysate powder described in Preparation Example 6 is performed
to produce lactoferrin hydrolysate powder.
[0158] Desalted cow's milk whey protein powder (produced by Milei
GmbH, 10 kg), milk casein powder (produced by Fonterra Limited, 6
kg), lactose (produced by Milei GmbH, 48 kg), mineral mixture
(produced by Tomita Pharmaceutical Co., Ltd., 920 g), vitamin
mixture (produced by Tanabe Seiyaku Co., Ltd., 32 g), the obtained
lactoferrin hydrolysate powder (17 to 690 g), and human milk
oligosaccharides (68 to 1698 g) are dissolved in warm water (300
kg), the solution is heated at 90.degree. C. for 10 minutes to
further dissolve the ingredients; modified fat (produced by TAIYO
YUSHI CORP., 28 kg) is added to the solution, and the mixture is
homogenized. Then, the homogenized mixture is subjected to
sterilization and concentration steps, and spray-dried to prepare a
nutritional composition containing lactoferrin hydrolysate and
human milk oligosaccharides (about 95 kg). If the obtained
nutritional composition is dissolved in water to prepare liquid
formula having a total solid concentration of 14% (w/v), which is
the standard solid concentration of formula milk, the lactoferrin
hydrolysate concentration and human milk oligosaccharide
concentration in the formula milk liquid will be 0.025 to 1 mg/mL
and 0.1 to 5 mg/mL, respectively.
[0159] By administering the nutritional composition produced as
described above to an infant or young child, viral infection of the
infant or young children can be prevented.
* * * * *